Nesta DIYbio

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Nesta DIYbio

  1. 1. Biohacking! (AKA Synthetic Biology for Beginners)Sunday, 10 February 13
  2. 2. Sunday, 10 February 13
  3. 3. Sunday, 10 February 13
  4. 4. Sunday, 10 February 13
  5. 5. Sunday, 10 February 13
  6. 6. Sunday, 10 February 13
  7. 7. Sunday, 10 February 13
  8. 8. Sunday, 10 February 13
  9. 9. Lab Rules • No food & drink in the lab • Gloves & aprons at all times • Wash/gel your hands when you leave the lab • Aargh spills! • No running, diving or bombingSunday, 10 February 13
  10. 10. Aseptic Technique • Keep air exposure to a minimum. • Don’t cross-contaminate.Sunday, 10 February 13
  11. 11. Micropipetting • Two different types • Set the volume by turning the dial • Suck! • Squirt! • Remember: Always use a fresh tip.Sunday, 10 February 13
  12. 12. …Sunday, 10 February 13
  13. 13. 1. • Iron Microbeads! (In tube 1) • Wash them… 50µl of washing buffer. Mix. • Clean them… pull the beads to one side with the magnet; extract the liquid. • Wash and clean them again…Sunday, 10 February 13
  14. 14. 2. • Ligation! Glues the first “part” to the beads. • Add 7µl of ligation mix (tube with the spot on) and mix. • Transfer this to the tube containing our first part (Tube 2) • Wait 5-10 minutes…Sunday, 10 February 13
  15. 15. 3. • Wash them… 50µl of washing buffer. Mix. • Clean them… pull them to one side with the magnet; extract the liquid. • Wash and clean them again…Sunday, 10 February 13
  16. 16. 4. • Ligation again! Glue the final two parts to our beads. • Add 7µl of ligation mix (tube with the spot on) and mix. • Transfer this to the tube containing our second part (Tube 3) and mix. • Transfer this to the tube containing our first part (Tube 4) and mix some more. • Wait 5-10 minutes…Sunday, 10 February 13
  17. 17. 5. • Wash them… 50µl of washing buffer. Mix. • Clean them… pull them to one side with the magnet; extract the liquid. • Wash and clean them again…Sunday, 10 February 13
  18. 18. 5. • Elution! Gets rid of the beads • Add 20µl of elution buffer and mix • Pull the beads to one side with the magnet • Extract the finished part in liquid form; put it into one of the empty tubes.Sunday, 10 February 13
  19. 19. …Sunday, 10 February 13
  20. 20. Modify that e.coli! • Put 100µl of Transformation buffer in an empty tube; put the tube on ice for 5 minutes, then… • Take a loop. Grab some e.coli and add it to the transformation buffer – “twiddle vigorously”. Wait for 10 minutes… • Add 40µl of your creation and mix. • Bring your tube to the front!Sunday, 10 February 13
  21. 21. Modify that e.coli! (Pt. 2) • “Heat shock” our e.coli for exactly 30 seconds in the water bath (42ºc) • Put them back on ice for 1-2 minutes • Add 100µl of recovery broth, keep them warm in your hand until…Sunday, 10 February 13
  22. 22. Modify that e.coli! (Pt. 3) • Put 100µl of your modified e.coli in a petri dish. • Remove the spreader from the alcohol and flame it (so there’s no alcohol left) • Spread the e.coli around your plate • …and that’s it! Bring your plate to the front and put it in the incubator.Sunday, 10 February 13
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